Overview | PrinterFriendlyVersion |
Ex/Em(nm) | 596/619 |
MW | ~800 |
CAS# | N/A |
Solvent | N/A |
Storage | F/D/L |
Category | CellBIOLOGy LabelingCells |
Related | FluorescenceImaging BiochemicalAssays |
Spectrum | AdvancedSpectrumViewer |
1.PrepareLysosome-stainingsolution:
1.1 WarmLysoBrite™dyestoroomtemperature.
1.2 Preparedyeworkingsolutionbydiluting20µLof500XLysoBrite™dyesto10mLofHanksand20mmHepesbufferorbuffer(HBSS)ofyourchoice.
Note1:20µLofLysoBrite™dyeisenoughforone96-wellplate.AliquotandstoreunusedLysoBrite™dyestocksolutionsat<-15oC.Protectitfromlightandavoidrepeatedfreeze-thawcycles.
Note2:Theoptimalconcentrationofthefluorescentlysosomeindicatorvariesdependingonthespecificapplication.Thestainingconditionsmaybemodifiedaccordingtotheparticularcelltypeandthepermeabilityofthecellsortissuestotheprobe.
2.Prepareandstaincells:
2.1 Foradherentcells:a).Growcellseitherina96-wellblackwall/clearbottomplate(100µL/well/96-wellplate)oroncover-slipsinsideapetridishfilledwiththeappropriateculturemedium.Whencellsreachthedesiredconfluence,addequalvolumeofthedye-workingsolution(fromStep1.2).b).Incubatethecellsina37°C,5%CO2incubatorfor30minutes.c).Washthecellstwicewithpre-warmed(37°C)Hanksand20mMHepesbuffer(HBSS)orbufferofyourchoice,fillthecellwellswithHBSSorgrowthmedium.d).Observethecellsusingafluorescencemicroscopefittedwithadesiredfilterset.
Note:Itisrecommendedtoincreaseeitherthelabelingconcentrationortheincubationtimetoallowthedyetoaccumulateifthecellsdonotappeartobesufficientlystained.
2.2 Forsuspensioncells:a).Addequalvolumeofdye-workingsolution(fromStep1.2)intothecells.Incubatethecellsina37°C,5%CO2incubatorfor30minutes.b).Washthecellstwicewithpre-warmed(37°C)Hanksand20mMHepesbuffer(HBSS)orbufferofyourchoice,fillthecellwellswithHBSSorgrowthmedium.c).Observethecellsusingafluorescencemicroscopeequippedwithadesiredfilterset.
Note1:Itisrecommendedtoincreaseeitherthelabelingconcentrationortheincubationtimetoallowthedyetoaccumulateifthecellsdonotappeartobesufficientlystained.
Note2:Suspensioncellsmaybeattachedtocover-slipsthathavebeentreatedwithBDCell-Tak®(BDBiosciences)andstainedasadherentcells(seeStep2.1).
References&Citations | CitationExplorer |
Autophagyproteinsarenotuniversallyrequiredforphagosomematuration
Authors:MarijaCemma,SergioGrinstein,JohnHBrumell
Journal:Autophagy(2016):1440--1446
Differentialdetectionoftumorcellsusingacombinationofcellrolling,multivalentbinding,andmultipleantibodies
Authors:JaHyeMyung,KhyatiAGajjar,JihuaChen,RobertEMolokie,SeungpyoHong
Journal:Analyticalchemistry(2014):6088--6094
Versatilefabricationofnanoscalesol--gelbioactiveglassparticlesforefficientbonetissueregeneration
Authors:BoLei,XiaofengChen,XueHan,JiaanZhou
Journal:JournalofMaterialsChemistry(2012):16906--16913
Advancedglycationend-productsincreaseIL-6andICAM-1expressionviaRAGE,MAPKandNF-$kappa$Bpathwaysinhumangingivalfibroblasts
Authors:KNonaka,YKajiura,MBando,ESakamoto,YInagaki,JHLew,KNaruishi,TIkuta,KYoshida,TKobayashi
Journal:JournalofPeriodontalResearch